Αρχειοθήκη ιστολογίου

Δευτέρα 18 Δεκεμβρίου 2017

Pharmacogenetics of KCNQ channel activation in 2 potassium channelopathy mouse models of epilepsy

Summary

Objectives

Antiseizure drugs are the leading therapeutic choice for treatment of epilepsy, but their efficacy is limited by pharmacoresistance and the occurrence of unwanted side effects. Here, we examined the therapeutic efficacy of KCNQ channel activation by retigabine in preventing seizures and neurocardiac dysfunction in 2 potassium channelopathy mouse models of epilepsy with differing severity that have been associated with increased risk of sudden unexpected death in epilepsy (SUDEP): the Kcna1−/− model of severe epilepsy and the Kcnq1A340E/A340E model of mild epilepsy.

Methods

A combination of behavioral, seizure threshold, electrophysiologic, and gene expression analyses was used to determine the effects of KCNQ activation in mice.

Results

Behaviorally, Kcna1−/− mice exhibited unexpected hyperexcitability instead of the expected sedative-like response. In flurothyl-induced seizure tests, KCNQ activation decreased seizure latency by ≥50% in Kcnq1 strain mice but had no effect in the Kcna1 strain, suggesting the influence of genetic background. However, in simultaneous electroencephalography and electrocardiography recordings, KCNQ activation significantly reduced spontaneous seizure frequency in Kcna1−/− mice by ~60%. In Kcnq1A340E/A340E mice, KCNQ activation produced adverse cardiac effects including profound bradycardia and abnormal increases in heart rate variability and atrioventricular conduction blocks. Analyses of Kcnq2 and Kcnq3 mRNA levels revealed significantly elevated Kcnq2 expression in Kcna1−/− brains, suggesting that drug target alterations may contribute to the altered drug responses.

Significance

This study shows that treatment strategies in channelopathy may have unexpected outcomes and that effective rebalancing of channel defects requires improved understanding of channel interactions at the circuit and tissue levels. The efficacy of KCNQ channel activation and manifestation of adverse effects were greatly affected by genetic background, potentially limiting KCNQ modulation as a way to prevent neurocardiac dysfunction in epilepsy and thereby SUDEP risk. Our data also uncover a potential role for KCNQ2-5 channels in autonomic control of chronotropy.



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Intraoral ultrasonography to measure tumor thickness of oral cancer: A systematic review and meta-analysis

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Publication date: February 2018
Source:Oral Oncology, Volume 77
Author(s): Thomas J.W. Klein Nulent, Rob Noorlag, Ellen M. Van Cann, Frank A. Pameijer, Stefan M. Willems, Adrian Yesuratnam, Antoine J.W.P. Rosenberg, Remco de Bree, Robert J.J. van Es
Early oral cancer is preferably treated by surgery. Its complete removal is essential for locoregional control and disease-free survival. Inadequate resection margins require adjuvant therapy such as re-resection or (chemo)radiation, that causes extra morbidity and oral discomfort. Intraoral ultrasonography (US) is reported to be of value in determining tumor thickness. Intraoperative visualization of the tumor may facilitate the resection and ensure adequate surgical margins. Furthermore, accurate prediction of tumor thickness could help determine the treatment strategy of the clinically node-negative neck, as thickness and depth of invasion are predictors of cervical metastasis as well as prognosticators of survival. The 8th edition of the American Joint Committee on Cancer staging system for oral squamous cell carcinoma has included depth of invasion as parameter for cT-stage. The aim of this review is to analyze the accuracy of intraoral US in determining tumor thickness in oral cancer.A systematic search was conducted, and the quality of the included papers was assessed using the QUADAS-2 tool for diagnostic accuracy studies. Subsequently, a meta-analysis was performed on the available individual participant data of 240 patients.Most of the twelve included studies focused on T1-2 tongue cancer (n = 129). Meta-analysis showed a high correlation in tumor thickness within this subgroup as measured by intraoral US and histopathology (r = 0.82, p < .001), with minor overestimation of 0.5 mm on US. It is concluded that intraoral US is very accurate in determining tumor thickness in early oral tongue cancer.



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Postoperative staging of the neck dissection using extracapsular spread and lymph node ratio as prognostic factors in HPV-negative head and neck squamous cell carcinoma patients

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Publication date: February 2018
Source:Oral Oncology, Volume 77
Author(s): Katarina Majercakova, Cristina Valero, Montserrat López, Jacinto García, Nuria Farré, Miquel Quer, Xavier León
ObjectivesThe presence of nodes with extracapsular spread (ECS) and the lymph node ratio (LNR) have prognostic competence in the pathologic evaluation of patients with a head and neck squamous cell carcinoma (HNSCC) treated with a neck dissection. The purpose of this study is to assess the effect of ECS & LNR on prognosis of HPV negative HNSCC patients treated with neck dissection and to compare to 8th edition TNM/AJCC classification.Materials and methodsWe carried out a retrospective study of 1383 patients with HNSCC treated with a neck dissection between 1985 and 2013. We developed a classification of the patients according to the presence of nodes with ECS and the LNR value with a recursive partitioning analysis (RPA) model.ResultsWe obtained a classification tree with four terminal nodes: for patients without ECS (including patients pN0) the cut-off point for LNR was 1.6%, while for patients with lymph nodes with ECS it was 11.4%. The 5-year disease-specific survival for patients without ECS/LNR < 1.6% was 83.3%; for patients without ECS/LNR ≥ 1.6% it was 61.5%; for patients with ECS/LNR < 11.4% it was 33.7%; and for patients with ECS/LNR ≥ 11.4% it was 18.5%. The classification obtained with RPA had better discrimination between categories than the 8th edition of the TNM/AJCC classification.ConclusionECS status and LNR value proved high prognostic capacity in the pathological evaluation of the neck dissection. The combination of ECS and LNR improved the predictive capacity of the 8th edition of the TNM/AJCC classification in HPV-negative HNSCC patients.



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Immolation dans notre centre de traitement des brûlés de 2011 à 2016

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Publication date: Available online 18 December 2017
Source:Annales de Chirurgie Plastique Esthétique
Author(s): S. Chatelain, K. Serror, M. Chaouat, M. Mimoun, D. Boccara
L'immolation par le feu est dans certains pays tels que l'Iran ou la Tunisie une pratique couramment employée afin de mettre fin à ses jours. Des études ont été réalisées sur ce sujet dans de nombreux pays développés et en voie de développement, permettant de faire ressortir des sujets à risque selon les pays. Cependant nous n'avons pas à ce jour de données récentes sur la population d'immolés par le feu en France. Nous avons réalisé une étude épidémiologique rétrospective sur les immolations entre 2011 et 2016 au centre de traitement des brûlés de l'hôpital Saint-Louis à Paris, en nous appuyant sur les facteurs favorisants retrouvés dans la littérature. Nous avons étudié la prévalence, et les caractéristiques des patients entrant en hospitalisation pour ce motif. Entre 2011 et 2016, 1098 patients ont été hospitalisés dans notre service. Cinquante patients ont été admis pour cause d'immolation, soit 5 % de la population hospitalisée. L'âge moyen à l'entrée était de quarante-six ans, majoritairement des hommes (62 %). La surface cutanée totale brûlée moyenne était de 34,5 %, et la durée moyenne de séjour de cinquante-trois jours. Neuf décès sont survenus dans cette population (18 %), chez des patients des deux sexes, brûlés à 62,5 % en moyenne et d'un âge similaire à celui de notre cohorte. Il s'agissait d'un passage à l'acte suicidaire pour quarante-neuf patients, avec seulement un cas d'immolation criminelle. Des antécédents psychiatriques ont été retrouvés chez trente-cinq patients (70 %). Vingt et un d'entre eux avaient déjà tenté de se suicider précédemment (42 %). Neuf patients étaient alcooliques chroniques et quatre toxicomanes. Les motifs fréquents de passage à l'acte étaient une rupture sentimentale, un décès ou des problèmes financiers. Les produits utilisés étaient le white spirit, l'alcool à brûler et l'essence. La prévalence des immolations dans notre étude est très faible par rapport à celle retrouvée dans les pays en voie de développement. La majorité de notre cohorte est composée de patients ayant des antécédents de troubles de l'humeur. Les antécédents de tentative de suicide sont également un facteur de risque majeur de passage à l'acte suicidaire par immolation. Des mesures de prévention devraient être mises en place pour prévenir ce risque.The prevalence of immolation by fire in France is uncertain. We carried out a retrospective epidemiological study on immolations between 2011 and 2016 at the burn treatment centre at the hôpital Saint-Louis in Paris. We studied the prevalence and characteristics of patients entering hospital for this reason. Between 2011 and 2016, a total of 1098 patients were hospitalized in the centre, of which 50 were admitted for immolation, i.e. five percent of the hospitalized population. The average age at entry was 46 years, and they were mostly men (62%). All but one was a suicide attempt. The average total burn area was 34.5%, and the average length of stay in the centre was 53 days. The products used for the immolation were mostly white spirit, alcohol or gasoline. Nine (18%) out of the 50 patients died, burned at 62.5% on average. A psychiatric history was reported in 35 patients and 21 had previously attempted suicide; nine patients were chronic alcoholics and four were drug addicts. The most frequently reported reasons for the suicide attempt were sentimental breakdown, death of a relative or financial problem. The prevalence of immolation in our study is very low compared to that found in developing countries. The majority of our cohort is composed of patients with a history of psychiatric disorders. The history of attempted suicide is also a major risk factor for committing suicidal acts by immolation. Prevention measures should be implemented to reduce this risk.



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Endoscopic transpterygoid approach to a mass in a child

Publication date: Available online 18 December 2017
Source:International Journal of Pediatric Otorhinolaryngology
Author(s): Dong Hoon Lee, Hee Jo Baek, Tae Mi Yoon, Joon Kyoo Lee, Sang Chul Lim
The endoscopic transterygoid approach to the petrous apex is a feasible/alternative approach in carefully selected patients with specific favorable anatomy, even children. This approach, unlike traditional approaches, spares cochlear and vestibular function. We report a case of a six-year-old boy with embryonal rhabdomyosarcoma of the petrous apex that was diagnosed via the endoscopic transpterygoid approach.



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Issue Cover (December 2017)

Thumbnail image of graphical abstract

Cover image by Dr. Natalie Doig (MRC Brain Network Dynamics Unit, Department of Pharmacology, Oxford). The cover image is of a frontal section of mouse brain showing many regions of the basal ganglia. The section was triple-immunostained to reveal tyrosine hydroxylase (TH; cyan), parvalbumin (PV; green) and choline acetyltransferase (magenta).



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Acknowledgment to reviewers 1st October 2016–30th September 2017



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SOCIETY NEWS



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A 72-Year-Old Male with A Slow Growing Pineal Region Tumor



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A 60-Year-Old Woman with Multifocal Subcortical Infarcts



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A 68-Year-Old Woman with A Left Orbital and Temporal Mass



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An 8-Year-Old Girl with A Supratentorial Mass



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A 39-Year-Old Woman with Progressive Vision Impairment



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A 44-Year-Old Female with Familial Mediterranean Fever, Cardiomyopathy and End Stage Renal Disease



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A 62-Year-Old Woman with A History of Muscle Pain and Skin Rash for 1 Month



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A 31-Year-Old Man with Slowly Progressive Limb Muscle Weakness and Respiratory Insufficiency



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Issue Information



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Alterations in comprehensive geriatric assessment decrease survival of elderly patients with cancer

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Publication date: February 2018
Source:European Journal of Cancer, Volume 90
Author(s): M. Frasca, P. Soubeyran, C. Bellera, M. Rainfray, K. Leffondre, S. Mathoulin-Pélissier
IntroductionA comprehensive geriatric assessment (CGA) evaluating several domains of health is recommended for elderly patients with cancer. Effects of altered domains on the risk of death in this population need to be clarified. The aim of this study was to estimate the independent association of each CGA domain to overall survival (OS).MethodPatients included in the ONCODAGE cohort completed a CGA at baseline. Cox models (one per domain) estimated the hazard ratio (HR) of death for each CGA domain. Directed Acyclic Graphs (DAGs) selected specific sets of adjustment factors for each model.ResultsThe analysis included 1264 patients (mean age: 78 years, women: 70%). Median follow-up was 5.2 years, and 446 patients died. Each altered domain had a detrimental effect on survival, sometimes dependent on gender, age, education or time from inclusion. Nutritional status had a time-varying effect, with higher mortality rates if altered only within the first 3 years of follow-up. In case of altered mobility, the risk of death was higher only for the youngest patients and, in case of altered autonomy, only for the youngest women. An altered neurological state led to higher mortality rates; this effect increased with the level of education. Patients with altered psychological status or more than four comorbidities at baseline had also higher mortality rates.ConclusionsPatients with an altered CGA domain have a higher risk of death than those without any alteration. The effect of some alterations is different in some subgroups or at a given time of the treatments.



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A randomised phase II trial of docetaxel versus docetaxel plus carboplatin in patients with castration-resistant prostate cancer who have progressed after response to prior docetaxel chemotherapy: The RECARDO trial

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Publication date: February 2018
Source:European Journal of Cancer, Volume 90
Author(s): Esther W. Bouman-Wammes, H. Pieter van den Berg, Linda de Munck, Aart Beeker, Carolien H. Smorenburg, Walter L. Vervenne, Juleon L.L.M. Coenen, Henk M.W. Verheul, Winald R. Gerritsen, Alfons J.M. Van den Eertwegh
BackgroundDocetaxel is standard first-line chemotherapy for patients with metastatic castration–resistant prostate carcinoma (mCRPC). Docetaxel re-challenge has never been tested in a prospective randomised controlled study. As some studies support the addition of carboplatin to docetaxel, we performed a phase II trial investigating the combination of docetaxel plus carboplatin versus docetaxel re-treatment in docetaxel pre-treated mCRPC patients.MethodsPatients with mCRPC with a progression-free interval of ≥3 months after initial docetaxel treatment were randomised between docetaxel 75 mg/m2 or docetaxel 60 mg/m2 plus carboplatin AUC4. The primary end-point was progression-free survival (PFS; PSA/RECIST).ResultsOwing to insufficient recruitment, the study was discontinued early after inclusion of 75 patients (targeted 150) PFS and overall survival (OS) were comparable between both groups (median PFS 12.7 months (95% CI 9.9–17.5 months) with docetaxel monotherapy and 11.7 months (95% CI 8.5–21.0 months) with combination therapy (p = 0.98); OS 18.5 months (95% CI 11.8–24.5 months) versus 18.9 months (95% CI 16.0–23.7 months) (p = 0.79). An interim analysis (SEQTEST) showed that the null hypothesis could already be excepted, and no significant difference between both study arms was expected if inclusion would be completed. The incidence of grade 3–4 infections and gastrointestinal side-effects was numerical higher in the carboplatin arm (p = 0.056).ConclusionThis early terminated study suggests no benefit from the addition of carboplatin to docetaxel re-treatment in patients with mCRPC, whereas the combination resulted in more toxicity. Re-treatment with docetaxel monotherapy appears to be feasible, save and effective for patients with mCRPC and an initial good response to docetaxel.Trial registrationNTR3070.



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Key High Efficiency Practices of Emergency Department Providers: A Mixed Methods Study

Abstract

Objective

The objective of this study was to determine specific provider practices associated with high provider efficiency in community emergency departments (EDs).

Methods

A mixed methods study design was utilized to identify key behaviors associated with efficiency:

Stage 1. A convenience sample of sixteen participants (ED medical directors, nurses, advanced practice providers, and physicians) identified provider efficiency behaviors during semi-structured interviews. Ninety-nine behaviors were identified and distilled by a group of three ED clinicians into 18 themes.

Stage 2. An observational study of 35 providers was performed in four (30,000 to 55,000-visit) community EDs during two 4-hour periods and recorded in minute-by-minute observation logs.

Stage 3. Each behavior or practice from Stage 1 was assigned a score within each observation period. Behaviors were tested for association with provider efficiency (Relative Value Units [RVUs] per hour) using linear univariate generalized estimating equations (GEE) with an identity link, clustered on ED site.

Results

Five ED provider practices were found to be positively associated with efficiency: average patient load, using name of team member, conversations with health care team, visits to patient rooms, and running the board. Two behaviors, "inefficiency practices", demonstrated significant negative correlations: non-work-related tasks and documentation on patients no longer in the ED.

Conclusions

Average patient load, running the board, conversations with team member, and using names of team members is associated with enhanced provider productivity. Identification of behaviors associated with efficiency can be utilized by medical directors, clinicians, and trainees to improve personal efficiency or counsel team members.

This article is protected by copyright. All rights reserved.



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Clinical examination for acute aortic dissection: A systematic review and meta-analysis

Abstract

Introduction

Acute aortic dissection is a life-threatening condition due to a tear in the aortic wall. It is difficult to diagnose and if missed carries a significant mortality.

Methods

We conducted a librarian assisted systematic review of Pubmed, Medline, Embase and the Cochrane database from 1968 to July 2016. Titles and abstracts were reviewed and data extracted by two independent reviewers (agreement measured by Kappa). Studies were combined if low clinical and statistical heterogeonity (I2<30%). Study quality was assessed using the QUADAS-2 tool. Bivariate random effects meta analyses using Revman 5 and SAS 9.3 were performed.

Results

We identified 792 records: 60 were selected for full text review, 9 studies with 2,400 participants were included (QUADAS-2 low risk of bias, Kappa 0.89(for full text review)). Prevalence of aortic dissection ranged from 21.9-76.1% (mean 39.1% SD 17.1%). The clinical findings increasing probability of aortic dissection were: 1) neurological deficit (n=3, specificity 95%, LR+ 4.4 95% CI 3.3-5.7, I2 0%), 2) hypotension (n=4, specificity 95%, LR+ 2.9 95% CI 1.8-4.6, I2 42%), and decreasing probability: absence of a widened mediastinum (n=4, sensitivity 76%-95%, LR- 0.14-0.60, I2 93%) and an American heart association (AHA) aortic dissection detection (ADD) risk score <1 (n=1 sensitivity 91%, LR- 0.22 95% CI 0.15–0.33).

Conclusions

Suspicion for acute aortic dissection should be raised with hypotension, pulse or neurological deficit. Conversely, a low AHA ADD score decreases suspicion. Clinical gestalt informed by high and low risk features together with an absence of an alternative diagnosis should drive investigation for acute aortic dissection.

This article is protected by copyright. All rights reserved.



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Post-surgical effects on language in patients with presumed low-grade glioma

Objectives

Low-grade glioma (LGG) is a slow-growing brain tumour often situated in or near areas involved in language and/or cognitive functions. Thus, language impairments due to tumour growth or surgical resection are obvious risks. We aimed to investigate language outcome following surgery in patients with presumed LGG, using a comprehensive and sensitive language assessment.

Materials and methods

Thirty-two consecutive patients with presumed LGG were assessed preoperative, early post-operative, and 3 months post-operative using sensitive tests including lexical retrieval, language comprehension and high-level language. The patients' preoperative language ability was compared with a reference group, but also with performance at post-operative controls. Further, the association between tumour location and language performance pre- and post-operatively was explored.

Results

Before surgery, the patients with presumed LGG performed worse on tests of lexical retrieval when compared to a reference group (BNT: LGG-group median 52, Reference-group median 54, = .002; Animals: LGG-group mean 21.0, Reference-group mean 25, P = 001; Verbs: LGG-group mean 17.3, Reference-group mean 21.4, = .001). At early post-operative assessment, we observed a decline in all language tests, whereas at 3 months there was only a decline on a single test of lexical retrieval (Animals: preoperative. median 20, post-op median 14, = .001).

The highest proportion of language impairment was found in the group with a tumour in language-eloquent areas at all time-points.

Conclusions

Although many patients with a tumour in the left hemisphere deteriorated in their language function directly after surgery, their prognosis for recovery was good.



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Is carotid imaging underused in patients with transient ischemic attack or ischemic stroke? A Swedish Stroke Register (Riksstroke) study

Background and aim

Carotid artery stenosis is one of the major causes of transient ischemic attack (TIA) and acute ischemic stroke (IS), and carotid surgery and stenting are used to reduce the risk of ipsilateral IS. However, the adherence to the recommendation of carotid imaging in clinical practice has not been well studied. We analyzed proportions of carotid imaging and determinants for its non-use in patients with TIA and IS with respect to baseline demographics, risk factors, hospital characteristics, and geographical region.

Patients and methods

Hospital-based data on TIA and IS events, registered from July 2011 to June 2013, were obtained from the Swedish Stroke Register (Riksstroke). Carotid imaging diagnostics included carotid Doppler ultrasound and computed tomography angiography.

Results

Carotid imaging was performed in 70% (10 545/15 021) of patients with TIA and 54% (23 772/44 075) of patients with IS. The most significant independent determinants for not undergoing carotid imaging were, in patients with TIA: age ≥85 year (odds ratio (OR), 7.3; 95% confidence interval (CI), 6.4-8.4) and a history of stroke (OR, 2.3; 95% CI, 2.1-2.5); and in patients with IS: age ≥85 year (OR, 9.8; 95% CI, 9.0-10.6), age 75-84 year (OR, 2.5; 95% CI, 2.3-2.7), and reduced level of consciousness at admission (OR, 3.4; 95% CI, 3.1-3.6). Care at a University hospital and in a stroke unit increased the likelihood of carotid imaging. There were substantial regional variations regarding proportions of carotid imaging.

Conclusion

Carotid imaging appears to be underused in patients with TIA and IS. Opportunities of secondary stroke prevention with carotid interventions are likely missed.



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Timing of tracheostomy in patients with prolonged endotracheal intubation: a systematic review

Abstract

The objective of this article is to evaluate the appropriate timing of tracheostomy in patients with prolonged intubationregarding the incidence of hospital-acquired pneumonia, mortality, length of stay in intensive care unit (ICU) and duration of artificial ventilation. The study included published articles yielded by a search concerning timing of tracheostomy in adult and pediatric patients with prolonged intubation. The search was limited to articles published in English language in the last 30 years (between 1987 and 2017). For the 690 relevant articles, we applied our inclusion and exclusion criteria and only 43 articles were included. 41 studies in the adult age group including 222,501 patients and 2 studies in pediatric age group including 140 patients met our criteria. Studies in adult age group were divided into three groups according to the methodology of determining the cut off timing for early tracheostomy, they were divided into studies that considered early tracheostomy within the first 7, 14 or 21 days of endotracheal intubation, while in pediatric age group the cut off timing for early tracheostomy was within the first 7 days of endotracheal intubation. There was a significant difference in favor of early tracheostomy in adults' three groups and pediatric age group as early tracheostomy was superior regarding reduced duration of mechanical ventilation, with less mortality rates and less duration of stay in ICU. Regarding hospital-acquired pneumonia, it was significantly less in adult groups but with no significant difference in pediatric age group (3 patients out of 72 pediatric patient with early tracheostomy had pneumonia compared to 11 patients out of 68 with late tracheostomy). Studies defining early tracheostomy as that done within 7 days of intubation had better results than those defining early tracheostomy as that done within 14 or 21 days of intubation. In conclusion, early tracheostomy within 7 days of intubation should be done for both adults and pediatric patients with prolonged intubation.



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Concussion Alters the Functional Brain Processes of Visual Attention and Working Memory

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Journal of Neurotrauma , Vol. 0, No. 0.


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Transcranial Doppler Systolic Flow Index and ICP-Derived Cerebrovascular Reactivity Indices in Traumatic Brain Injury

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Journal of Neurotrauma , Vol. 0, No. 0.


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Acute Changes in Plasma Total Tau Levels Are Independent of Subconcussive Head Impacts in College Football Players

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Journal of Neurotrauma , Vol. 0, No. 0.


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Reduced Field of View Diffusion Tensor Imaging and Fiber Tractography of the Pediatric Cervical and Thoracic Spinal Cord Injury

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Journal of Neurotrauma , Vol. 0, No. 0.


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Post-Injury Administration of Galantamine Reduces Traumatic Brain Injury Pathology and Improves Outcome

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Journal of Neurotrauma , Vol. 0, No. 0.


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Incidence and Natural Progression of Neurogenic Shock after Traumatic Spinal Cord Injury

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Journal of Neurotrauma , Vol. 0, No. 0.


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Increased infiltration of CD11c+/CD123+ dendritic cell subsets and upregulation of TLR/IFN-α signaling participate in pathogenesis of oral lichen planus

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Publication date: Available online 18 December 2017
Source:Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology
Author(s): Yufeng Wang, Shu Shang, Qianqian Sun, Junjun Chen, Guanhuan Du, Hong Nie, Xiaozhe Han, Guoyao Tang
ObjectiveInvestigation of dendritic cell (DC) subsets and expression patterns of Toll-like receptors (TLRs) was conducted to understand the pathogenesis in oral lichen planus (OLP).Study DesignBlood, OLP lesion and control samples were collected. Four DC subsets (CD11c+CD123-myeloid DC-mDC1, CD141+myeloid DC-mDC2, CD11c-CD123+plasmacytoid DC-pDC and CD1a+CD207+Langerhans cells-LC) were investigated via flow cytometry (FCM) and immunohistochemical (IHC) staining. Expression patterns of TLRs and their downstream molecules were analyzed via qRT-PCR and IHC in situ.ResultsThirty-two samples were collected (9 controls and 23 OLP patients). FCM results showed the percentages of LC, mDC1, mDC2 and pDC in situ were 0.0119±0.0251%, 0.0064±0.0134%, 0.0005±0.0011%, and 0.0022±0.0019% in control mucosa, respectively. The mDC1 (0.0300±0.0276%) and pDC (0.0204±0.0186%) subsets were significantly increased in OLP lesions (p<0.01). No marked differences were evident, when comparing all four DC subsets from blood, between control and OLP groups. Significant upregulation of TLR7, TLR8 and TLR9 were disclosed in OLP (p<0.01), along with their downstream interferon-α(IFN-α) signaling molecules(IRF7 and IFN-α, p<0.01).ConclusionOur findings of increased infiltration of pDC and mDC1, along with upregulation of TLR/IFN-α signaling, provide valuable information for further understanding the immunity in OLP.



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An actionable test using loss of heterozygosity in identifying high-risk oral premalignant lesions

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Publication date: Available online 18 December 2017
Source:Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology
Author(s): Kelly Y.P. Liu, X.J. David Lu, Yi-Shing L. Cheng, Hagen Klieb, Samson Ng, Kelly McNeil, Aly Karsan, Catherine F. Poh
Objectives.To develop an actionable test using fluorescent-labeled primers with capillary gel electrophoresis (FCE) to assess loss of heterozygosity (LOH) of histologically similar low-grade lesions (LGLs) to identify high-risk lesions for oral cancer progression.Study Design.To determine the cut-offs of LOH, the FCE results of 52 surgical margin samples were used to compare to the existing LOH results from the previously validated 32P-GE approach. Using the developed FCE workflow, an independent set of 102 LGLs with known progression status was used to determine the LOH molecular risk (MR) patterns and associated risk of progression.Results.Using 65% cut-off LOH-FCE, the agreement of LOH-32P-GE showed an average of 82.3% (76.8-87.8). Comparing to non-progressors (n=61), anatomical site, and MR patterns (LOH at 9p21, 3p14, or 17p13) were independent risk factors. High-risk profile of tongue and MR3 (LOH at 9p21 and/or 3p14, and 17p13) was significantly associated with progression (HR, 6.7; 95% CI, 2.6-17.6) with specificity of 98.4% at identifying progressors.Conclusions.We have developed an objective, fast, environmental-safe, and cost-effective test using LOH to stratify the risk of LGLs. With further validation, it can be used in the clinical settings to provide clinicians additional information guiding the management of these lesions.



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Timing of tracheostomy in patients with prolonged endotracheal intubation: a systematic review

Abstract

The objective of this article is to evaluate the appropriate timing of tracheostomy in patients with prolonged intubationregarding the incidence of hospital-acquired pneumonia, mortality, length of stay in intensive care unit (ICU) and duration of artificial ventilation. The study included published articles yielded by a search concerning timing of tracheostomy in adult and pediatric patients with prolonged intubation. The search was limited to articles published in English language in the last 30 years (between 1987 and 2017). For the 690 relevant articles, we applied our inclusion and exclusion criteria and only 43 articles were included. 41 studies in the adult age group including 222,501 patients and 2 studies in pediatric age group including 140 patients met our criteria. Studies in adult age group were divided into three groups according to the methodology of determining the cut off timing for early tracheostomy, they were divided into studies that considered early tracheostomy within the first 7, 14 or 21 days of endotracheal intubation, while in pediatric age group the cut off timing for early tracheostomy was within the first 7 days of endotracheal intubation. There was a significant difference in favor of early tracheostomy in adults' three groups and pediatric age group as early tracheostomy was superior regarding reduced duration of mechanical ventilation, with less mortality rates and less duration of stay in ICU. Regarding hospital-acquired pneumonia, it was significantly less in adult groups but with no significant difference in pediatric age group (3 patients out of 72 pediatric patient with early tracheostomy had pneumonia compared to 11 patients out of 68 with late tracheostomy). Studies defining early tracheostomy as that done within 7 days of intubation had better results than those defining early tracheostomy as that done within 14 or 21 days of intubation. In conclusion, early tracheostomy within 7 days of intubation should be done for both adults and pediatric patients with prolonged intubation.



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Tinea capitis mimicking dissecting cellulitis in three children

Abstract

Tinea capitis mimicking dissecting cellulitis is a rare presentation, and there is a paucity of information regarding this presentation in the literature. Three children 10-14 years of age who presented with an unusual clinical manifestation of tinea capitis that clinically resembled dissecting cellulitis are reported. The patients were treated with systemic antifungals for 3-4 months. Treatment success was measured according to repeat fungal cultures and clinical assessment of hair regrowth at follow-up visits. All three patients had resolution of infection, with negative repeat fungal cultures and complete hair regrowth without scarring. These cases highlight a rare inflammatory subtype of tinea capitis that can be easily misdiagnosed and therefore improperly treated, prolonging the duration of infection.



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Editorial



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Referees



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Issue Information



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Building a state space for song learning

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Publication date: April 2018
Source:Current Opinion in Neurobiology, Volume 49
Author(s): Emily Lambert Mackevicius, Michale Sean Fee
The songbird system has shed light on how the brain produces precisely timed behavioral sequences, and how the brain implements reinforcement learning (RL). RL is a powerful strategy for learning what action to produce in each state, but requires a unique representation of the states involved in the task. Songbird RL circuitry is thought to operate using a representation of each moment within song syllables, consistent with the sparse sequential bursting of neurons in premotor cortical nucleus HVC. However, such sparse sequences are not present in very young birds, which sing highly variable syllables of random lengths. Here, we review and expand upon a model for how the songbird brain could construct latent sequences to support RL, in light of new data elucidating connections between HVC and auditory cortical areas. We hypothesize that learning occurs via four distinct plasticity processes: 1) formation of 'tutor memory' sequences in auditory areas; 2) formation of appropriately-timed latent HVC sequences, seeded by inputs from auditory areas spontaneously replaying the tutor song; 3) strengthening, during spontaneous replay, of connections from HVC to auditory neurons of corresponding timing in the 'tutor memory' sequence, aligning auditory and motor representations for subsequent song evaluation; and 4) strengthening of connections from premotor neurons to motor output neurons that produce the desired sounds, via well-described song RL circuitry.



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Modeling pathogenesis and treatment response in childhood absence epilepsy

Summary

Objective

Childhood absence epilepsy (CAE) is a genetic generalized epilepsy syndrome with polygenic inheritance, with genes for γ-aminobutyric acid (GABA) receptors and T-type calcium channels implicated in the disorder. Previous studies of T-type calcium channel electrophysiology have shown genetic changes and medications have multiple effects. The aim of this study was to use an established thalamocortical computer model to determine how T-type calcium channels work in concert with cortical excitability to contribute to pathogenesis and treatment response in CAE.

Methods

The model is comprised of cortical pyramidal, cortical inhibitory, thalamocortical relay, and thalamic reticular single-compartment neurons, implemented with Hodgkin-Huxley model ion channels and connected by AMPA, GABAA, and GABAB synapses. Network behavior was simulated for different combinations of T-type calcium channel conductance, inactivation time, steady state activation/inactivation shift, and cortical GABAA conductance.

Results

Decreasing cortical GABAA conductance and increasing T-type calcium channel conductance converted spindle to spike and wave oscillations; smaller changes were required if both were changed in concert. In contrast, left shift of steady state voltage activation/inactivation did not lead to spike and wave oscillations, whereas right shift reduced network propensity for oscillations of any type.

Significance

These results provide a window into mechanisms underlying polygenic inheritance in CAE, as well as a mechanism for treatment effects and failures mediated by these channels. Although the model is a simplification of the human thalamocortical network, it serves as a useful starting point for predicting the implications of ion channel electrophysiology in polygenic epilepsy such as CAE.



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Timing of tracheostomy in patients with prolonged endotracheal intubation: a systematic review

Abstract

The objective of this article is to evaluate the appropriate timing of tracheostomy in patients with prolonged intubationregarding the incidence of hospital-acquired pneumonia, mortality, length of stay in intensive care unit (ICU) and duration of artificial ventilation. The study included published articles yielded by a search concerning timing of tracheostomy in adult and pediatric patients with prolonged intubation. The search was limited to articles published in English language in the last 30 years (between 1987 and 2017). For the 690 relevant articles, we applied our inclusion and exclusion criteria and only 43 articles were included. 41 studies in the adult age group including 222,501 patients and 2 studies in pediatric age group including 140 patients met our criteria. Studies in adult age group were divided into three groups according to the methodology of determining the cut off timing for early tracheostomy, they were divided into studies that considered early tracheostomy within the first 7, 14 or 21 days of endotracheal intubation, while in pediatric age group the cut off timing for early tracheostomy was within the first 7 days of endotracheal intubation. There was a significant difference in favor of early tracheostomy in adults' three groups and pediatric age group as early tracheostomy was superior regarding reduced duration of mechanical ventilation, with less mortality rates and less duration of stay in ICU. Regarding hospital-acquired pneumonia, it was significantly less in adult groups but with no significant difference in pediatric age group (3 patients out of 72 pediatric patient with early tracheostomy had pneumonia compared to 11 patients out of 68 with late tracheostomy). Studies defining early tracheostomy as that done within 7 days of intubation had better results than those defining early tracheostomy as that done within 14 or 21 days of intubation. In conclusion, early tracheostomy within 7 days of intubation should be done for both adults and pediatric patients with prolonged intubation.



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Pleiotropic Roles for ZEB1 in Cancer

ZEB1 is a prime element of a network of transcription factors that controls epithelial-to-mesenchymal transition (EMT), a reversible embryonic transdifferentiation program that allows partial or complete transition from an epithelial to a mesenchymal state. Aberrant expression of ZEB1 has been reported in a variety of human cancers, where it is generally believed to foster migration, invasion, and metastasis. Over the past few years, in vitro and in vivo observations have highlighted unsuspected intrinsic oncogenic functions of ZEB1 that impact tumorigenesis from its earliest stages. Located downstream of regulatory processes that integrate microenvironmental signals and directly implicated in feedback loops controlled by miRNAs, ZEB1 appears to be a central switch that determines cell fate. Its expression fosters malignant transformation through the mitigation of critical oncosuppressive pathways and through the conferment of stemness properties. ZEB1 is also a key determinant of cell plasticity, endowing cells with the capacity to withstand an aberrant mitogenic activity, with a profound impact on the genetic history of tumorigenesis, and to adapt to the multiple constraints encountered over the course of tumor development. Cancer Res; 78(1); 1–6. ©2017 AACR.

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New Perspectives, Opportunities, and Challenges in Exploring the Human Protein Kinome

The human protein kinome comprises 535 proteins that, with the exception of approximately 50 pseudokinases, control intracellular signaling networks by catalyzing the phosphorylation of multiple protein substrates. While a major research focus of the last 30 years has been cancer-associated Tyr and Ser/Thr kinases, over 85% of the kinome has been identified to be dysregulated in at least one disease or developmental disorder. Despite this remarkable statistic, for the majority of protein kinases and pseudokinases, there are currently no inhibitors progressing toward the clinic, and in most cases, details of their physiologic and pathologic mechanisms remain at least partially obscure. By curating and annotating data from the literature and major public databases of phosphorylation sites, kinases, and disease associations, we generate an unbiased resource that highlights areas of unmet need within the kinome. We discuss strategies and challenges associated with characterizing catalytic and noncatalytic outputs in cells, and describe successes and new frontiers that will support more comprehensive cancer-targeting and therapeutic evaluation in the future. Cancer Res; 78(1); 1–15. ©2017 AACR.

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Novel Targeting of Transcription and Metabolism in Glioblastoma

Purpose: Glioblastoma (GBM) is highly resistant to treatment, largely due to disease heterogeneity and resistance mechanisms. We sought to investigate a promising drug that can inhibit multiple aspects of cancer cell survival mechanisms and become effective therapeutics for GBM patients. Experimental Design: To investigate TG02, an agent with known penetration of the Blood-Brain Barrier, we examined the effects as single agent and in combination with temozolomide, a commonly used chemotherapy in GBM. We utilized human GBM cells and a syngeneic mouse orthotopic GBM model, evaluating survival and the pharmacodynamics of TG02.  Mechanistic studies included TG02-induced transcriptional regulation, apoptosis and RNA sequencing in treated GBM cells as well as the investigation of mitochondrial and glycolytic function assays. Results: We demonstrated that TG02 inhibited cell proliferation, induced cell death, and synergized with temozolomide in GBM cells with different genetic background but not in astrocytes. TG02-induced cytotoxicity was blocked by the overexpression of phosphorylated CDK9, suggesting a CDK9-dependent cell killing. TG02 suppressed transcriptional progression of anti-apoptotic proteins, and induced apoptosis in GBM cells. We further demonstrated that TG02 caused mitochondrial dysfunction and glycolytic suppression and ultimately ATP depletion in GBM. A prolonged survival was observed in GBM mice receiving combined treatment of TG02 and temozolomide. The TG02-induced decrease of CDK9 phosphorylation was confirmed in the brain tumor tissue. Conclusions: TG02 inhibits multiple survival mechanisms and synergistically decreases energy production with temozolomide, representing a promising therapeutic strategy in GBM, currently under investigation in an ongoing clinical trial.



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Physical Education Classes, Physical Activity, and Sedentary Behavior in Children

ABSTRACTPurposeTo examine the associations between participation frequency in Physical Education (PE) classes and objective measures of physical activity (PA) and sedentary behavior (SB) in children from 12 countries at different levels of development.MethodsThis multinational, cross-sectional study included 5,874 children aged 9-11 years from sites in Australia, Brazil, Canada, China, Colombia, Finland, India, Kenya, Portugal, South Africa, the United Kingdom and the United States. PA and SB were monitored over 7 consecutive days using a waist-worn accelerometer. PA and SB data were presented for weekdays (times in- and out-of-school) and weekend days. Participation frequency in PE classes was determined by questionnaire. Multilevel modeling analyses stratified by sex were used.ResultsOverall, 24.8% of children self-reported participation in PE classes ≥ 3 times/week (25.3% in high-income countries [HIC], and 24.3% in low- and middle-income countries [LMIC]). After adjusting for age, sex, parental education and body mass index z-score, results showed that children from LMIC who took PE classes 1-2 times/week were more likely to present better indicators of PA and shorter time in SB in- and out-of-school. In HIC, boys that participated in PE classes were more likely to meet the moderate-to-vigorous PA (MVPA) recommendations and to present better indicators of PA (in school) and shorter time in SB in- and out-of-school. For girls in HIC, attending PE classes increased the likelihood of spending more time in MVPA, especially if they attended ≥ 3 times/week.ConclusionAttending PE classes is associated with a higher level of PA and lower level of SB in- and out-of-school during weekdays in children from countries at various levels of development. Purpose To examine the associations between participation frequency in Physical Education (PE) classes and objective measures of physical activity (PA) and sedentary behavior (SB) in children from 12 countries at different levels of development. Methods This multinational, cross-sectional study included 5,874 children aged 9-11 years from sites in Australia, Brazil, Canada, China, Colombia, Finland, India, Kenya, Portugal, South Africa, the United Kingdom and the United States. PA and SB were monitored over 7 consecutive days using a waist-worn accelerometer. PA and SB data were presented for weekdays (times in- and out-of-school) and weekend days. Participation frequency in PE classes was determined by questionnaire. Multilevel modeling analyses stratified by sex were used. Results Overall, 24.8% of children self-reported participation in PE classes ≥ 3 times/week (25.3% in high-income countries [HIC], and 24.3% in low- and middle-income countries [LMIC]). After adjusting for age, sex, parental education and body mass index z-score, results showed that children from LMIC who took PE classes 1-2 times/week were more likely to present better indicators of PA and shorter time in SB in- and out-of-school. In HIC, boys that participated in PE classes were more likely to meet the moderate-to-vigorous PA (MVPA) recommendations and to present better indicators of PA (in school) and shorter time in SB in- and out-of-school. For girls in HIC, attending PE classes increased the likelihood of spending more time in MVPA, especially if they attended ≥ 3 times/week. Conclusion Attending PE classes is associated with a higher level of PA and lower level of SB in- and out-of-school during weekdays in children from countries at various levels of development. *Corresponding Author: Mark S. Tremblay, Healthy Active Living and Obesity Research Group, Children's Hospital of Eastern Ontario Research Institute, 401 Smyth Road, Ottawa, Ontario, Canada, K1H 8L1. Phone: +1 613 737 7600 ext. 4114. Fax: +1 613 738 4800. E-mail: mtremblay@cheo.on.ca The International Study of Childhood Obesity, Lifestyle, and the Environment (ISCOLE) was funded by The Coca-Cola Company. GH was supported by a grant from the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health under Award Number R01DK100790. The funder had no role in the design and conduct of the study; collection, management, analysis and interpretation of the data; and preparation, review, or approval of the manuscript. The authors declare no conflicts of interest.The results of the present study do not constitute endorsement by the American College of Sports Medicine. The authors declare that the results of the study are presented clearly, honestly, and without fabrication, falsification, or inappropriate data manipulation. Accepted for Publication: 8 December 2017 © 2017 American College of Sports Medicine

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Lower Extremity Injury Increases Risk of First-time Low Back Pain in the U.S. Army

ABSTRACTLow back pain (LBP) and lower extremity injuries (LEI) are primary reasons for lost duty days and disability among military populations.PURPOSEThis study examined acute LEI as a risk factor for developing LBP and examined the time to incident LBP between individuals with and without a history of LEI.METHODSThis retrospective cohort study examined U.S. Army medical and personnel data from the Total Army Injury and Health Outcomes Database (TAIHOD) for the years 2007-2011. Anderson-Gill Cox Regression methods were used to examine the change in LEI status over time and changes in demographic covariates. Adjusted hazard ratios (HR) for LBP following LEI were calculated from the Cox regression model for each calendar year. An accelerated failure time (AFT) model was used to describe time to LBP, and mean time to event and adjusted time ratios (TR) following LEI were calculated from the AFT model for each year. Overall HR and TR for LBP following LEI were calculated over the 5 calendar years using variance-based weighted averages.RESULTSEach yearly analysis included an average of 213,307 Soldiers; on average for each year 8.44% of Soldiers developed LBP and 11.54% had previous LEI. The pooled time ratio showed Soldiers with a LEI had a 10% decrease in mean survival times to LBP compared to those without a LEI [TR=0.901, 95%CI (0.897, 0.905)]. The weighted average HR showed that Soldiers with a LEI had 1.7 times the hazard of LBP compared to those without LEI [HR=1.70, 95%CI (1.66, 1.74)].CONCLUSIONThese findings suggest that a potential second order effect of LEI is an increased short-term risk for developing LBP, which should be considered during rehabilitation planning. Low back pain (LBP) and lower extremity injuries (LEI) are primary reasons for lost duty days and disability among military populations. PURPOSE This study examined acute LEI as a risk factor for developing LBP and examined the time to incident LBP between individuals with and without a history of LEI. METHODS This retrospective cohort study examined U.S. Army medical and personnel data from the Total Army Injury and Health Outcomes Database (TAIHOD) for the years 2007-2011. Anderson-Gill Cox Regression methods were used to examine the change in LEI status over time and changes in demographic covariates. Adjusted hazard ratios (HR) for LBP following LEI were calculated from the Cox regression model for each calendar year. An accelerated failure time (AFT) model was used to describe time to LBP, and mean time to event and adjusted time ratios (TR) following LEI were calculated from the AFT model for each year. Overall HR and TR for LBP following LEI were calculated over the 5 calendar years using variance-based weighted averages. RESULTS Each yearly analysis included an average of 213,307 Soldiers; on average for each year 8.44% of Soldiers developed LBP and 11.54% had previous LEI. The pooled time ratio showed Soldiers with a LEI had a 10% decrease in mean survival times to LBP compared to those without a LEI [TR=0.901, 95%CI (0.897, 0.905)]. The weighted average HR showed that Soldiers with a LEI had 1.7 times the hazard of LBP compared to those without LEI [HR=1.70, 95%CI (1.66, 1.74)]. CONCLUSION These findings suggest that a potential second order effect of LEI is an increased short-term risk for developing LBP, which should be considered during rehabilitation planning. Corresponding Author: Joseph F. Seay, Ph.D., Military Performance Division, United States Army Research Institute of Environmental Medicine, 10 General Greene Avenue, Building 42, Natick, MA 01760, 508.233.4888 (Phone), joseph.f.seay.civ@mail.mil The opinions or assertions contained herein are the private views of the author(s) and are not to be construed as official or as reflecting the views of the Army or the Department of Defense. The investigators have adhered to the policies for protection of human subjects as prescribed in Army Regulation 70-25, and the research was conducted in adherence with the provisions of 32 CFR Part 219. The authors have no conflicts of interest or financial disclosures to report. Results and conclusions of the study do not constitute endorsement of the American College of Sports Medicine. Accepted for Publication: 6 December 2017 © 2017 American College of Sports Medicine

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Differences in Hip and Knee Running Moments across Female Pubertal Development

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AbstractPurposeTo investigate whether knee and hip running moments differ across stages of female pubertal development.MethodsThis was a cross sectional study comparing the barefoot running moments of 91 pre-pubertal (n=31, Tanner stage I), early/mid-pubertal (n=30, Tanner stage II-III) and late/post-pubertal (n=30, Tanner stage IV-V) girls. External peak moments for knee abduction (KAbM), knee adduction (KAM), knee flexion (KFM) and knee internal rotation (KIRM) were analyzed. Secondary measures of hip adduction moment (HAM) at time of peak KAbM and hip flexion moment (HFM) at time of peak KFM were also derived. Between-group differences were analyzed using a series of one-way ANOVAs and ANCOVAs.ResultsAt the knee, the late/post-pubertal girls displayed a higher peak KFM and KAM compared to the pre-pubertal group (p0.05). At the hip, both the late/post- (p=0.03) and early/mid-pubertal girls (p= 0.039) ran with a lower HAM at time of peak KAbM than the pre-pubertal girls. The HFM at time of peak KFM in late/post-pubertal girls was also significantly lower than both the early/mid- and pre-pubertal girls. (p0.05). At the hip, both the late/post- (p=0.03) and early/mid-pubertal girls (p= 0.039) ran with a lower HAM at time of peak KAbM than the pre-pubertal girls. The HFM at time of peak KFM in late/post-pubertal girls was also significantly lower than both the early/mid- and pre-pubertal girls. (p

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Primary surgical management with radial forearm free flap reconstruction in T4 oropharyngeal cancer: Complications and functional outcomes

Functional outcomes and complication rates after open surgery for advanced-stage oropharyngeal cancers are rarely reported. These measures are critical for choice of treatment modality and patient counseling. We describe the long term functional outcomes and associated complications of primary surgical management of T4 oropharyngeal cancers reconstructed with radial forearm free flaps.

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Electrophysiological evidence of language switching for bidialectals: an event-related potential study

Language switching is an important issue in the study of bilinguals. However, how the nontarget language affects the production of the target language under the language switch condition is still unclear, especially for bilinguals who speak two dialects (bidialectals). In the present study, we investigate this issue by a picture-naming task and using an event-related potential (ERP) technique. Two groups of proficient bidialectals, Mandarin (L1)–Cantonese (L2) and Cantonese (L1)–Mandarin (L2), participated in the study. They were required to name pictures with Mandarin or Cantonese under language switching conditions and language nonswitching conditions. The results showed that participants of both groups showed significant longer reaction time, larger P200, and smaller N400 under language switching conditions than that under language nonswitching conditions, which reflects the language switching costs. Moreover, participants of the two groups showed different P200 and N400 between Mandarin–Cantonese (MC) switching conditions and Cantonese–Mandarin (CM) switching conditions. Specifically, MC bidialectals showed larger P200 under the CM condition than that under the MC condition, whereas CM bidialectals showed an opposite P200 pattern (CM conditionCM condition). These findings supported the language-unspecific selection theory. Overall, our study is the first to provide electrophysiological evidence of language switching between two dialects. Correspondence to Dr Zhuoming Chen, The First Affiliated Hospital of Jinan University, 601 Huangpu Road West, Guangzhou 510630, China Tel: +86 133 9269 2183; fax: +86 020 8521 8459; e-mail: tchzm@21cn.com Received October 12, 2017 Accepted November 21, 2017 © 2017 Wolters Kluwer Health | Lippincott Williams & Wilkins

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Pretreatment with minocycline improves neurogenesis and behavior performance after midazolam exposure in neonatal rats

Laboratory studies suggested that general anesthetics induce neuroapoptosis and inhibit neurogenesis in developing brains of animals. Minocycline exerts neuroprotection against a wide range of toxic insults in neurodegenerative diseases models. Here, we investigate whether minocycline can alleviate neurogenetic damage and improve cognition following midazolam exposure in neonatal rats. Postnatal 7 days rats were divided randomly into three groups: control group (C), midazolam group (M), and minocycline pretreatment group (MP). After exposure to midazolam, the cell proliferation in the subventricular zone (SVZ) and the subgranular zone (SGZ) of the hippocampus in pups was analyzed by bromodeoxyuridine immunochemistry at 7 days after the administration of anesthesia. Cognitive function was assessed using the Morris water-maze test at 35 days after midazolam exposure. Compared with the control, midazolam reduced cell proliferation both in the SVZ and in the SGZ of the hippocampus of neonatal rats, and decreased spatial learning and memory ability of rats in adulthood significantly. Pretreatment with minocycline increased cell proliferation both in the SVZ and in the SGZ of the hippocampus and improved spatial learning and memory ability compared with midazolam, but it did not mitigate the changes to the normal levels compared with the controls. Our results indicated that pretreatment with minocycline can alleviate midazolam-induced damage in neural stem cell proliferation of neonatal rats and improve spatial learning and memory ability of rats in adulthood. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. http://ift.tt/1hexVwJ * Praveen K. Giri and Yang Lu contributed equally to the writing of article. Correspondence to Pengbo Zhang, PhD, MD, Department of Anesthesiology, The Second Affiliated Hospital, Xi'an Jiaotong University, 157# West 5 Road, Xi'an, Shaanxi 710004, China Tel: +86 298 767 9635; fax: +86 298 767 8005; e-mail: zhpbo@mail.xjtu.edu.cn Received August 20, 2017 Accepted October 11, 2017 © 2017 Wolters Kluwer Health | Lippincott Williams & Wilkins

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Least Injurious Mechanical Ventilation in Pulmonary Resection Surgery

No abstract available

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To Clot or Not to Clot: Understanding Coagulopathy in Liver Disease

imageNo abstract available

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Unadjusted Bivariate Two-Group Comparisons: When Simpler is Better

imageHypothesis testing involves posing both a null hypothesis and an alternative hypothesis. This basic statistical tutorial discusses the appropriate use, including their so-called assumptions, of the common unadjusted bivariate tests for hypothesis testing and thus comparing study sample data for a difference or association. The appropriate choice of a statistical test is predicated on the type of data being analyzed and compared. The unpaired or independent samples t test is used to test the null hypothesis that the 2 population means are equal, thereby accepting the alternative hypothesis that the 2 population means are not equal. The unpaired t test is intended for comparing dependent continuous (interval or ratio) data from 2 study groups. A common mistake is to apply several unpaired t tests when comparing data from 3 or more study groups. In this situation, an analysis of variance with post hoc (posttest) intragroup comparisons should instead be applied. Another common mistake is to apply a series of unpaired t tests when comparing sequentially collected data from 2 study groups. In this situation, a repeated-measures analysis of variance, with tests for group-by-time interaction, and post hoc comparisons, as appropriate, should instead be applied in analyzing data from sequential collection points. The paired t test is used to assess the difference in the means of 2 study groups when the sample observations have been obtained in pairs, often before and after an intervention in each study subject. The Pearson chi-square test is widely used to test the null hypothesis that 2 unpaired categorical variables, each with 2 or more nominal levels (values), are independent of each other. When the null hypothesis is rejected, 1 concludes that there is a probable association between the 2 unpaired categorical variables. When comparing 2 groups on an ordinal or nonnormally distributed continuous outcome variable, the 2-sample t test is usually not appropriate. The Wilcoxon-Mann-Whitney test is instead preferred. When making paired comparisons on data that are ordinal, or continuous but nonnormally distributed, the Wilcoxon signed-rank test can be used. In analyzing their data, researchers should consider the continued merits of these simple yet equally valid unadjusted bivariate statistical tests. However, the appropriate use of an unadjusted bivariate test still requires a solid understanding of its utility, assumptions (requirements), and limitations. This understanding will mitigate the risk of misleading findings, interpretations, and conclusions.

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Anesthesia & Analgesia: Update and a Year in Review

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A Novel Method of Evaluating Key Factors for Success in a Multifaceted Critical Care Fellowship Using Data Envelopment Analysis

imageBACKGROUND: The current system of summative multi-rater evaluations and standardized tests to determine readiness to graduate from critical care fellowships has limitations. We sought to pilot the use of data envelopment analysis (DEA) to assess what aspects of the fellowship program contribute the most to an individual fellow's success. DEA is a nonparametric, operations research technique that uses linear programming to determine the technical efficiency of an entity based on its relative usage of resources in producing the outcome. DESIGN: Retrospective cohort study. SUBJECTS AND SETTING: Critical care fellows (n = 15) in an Accreditation Council for Graduate Medical Education (ACGME) accredited fellowship at a major academic medical center in the United States. METHODS: After obtaining institutional review board approval for this retrospective study, we analyzed the data of 15 anesthesiology critical care fellows from academic years 2013–2015. The input-oriented DEA model develops a composite score for each fellow based on multiple inputs and outputs. The inputs included the didactic sessions attended, the ratio of clinical duty works hours to the procedures performed (work intensity index), and the outputs were the Multidisciplinary Critical Care Knowledge Assessment Program (MCCKAP) score and summative evaluations of fellows. RESULTS: A DEA efficiency score that ranged from 0 to 1 was generated for each of the fellows. Five fellows were rated as DEA efficient, and 10 fellows were characterized in the DEA inefficient group. The model was able to forecast the level of effort needed for each inefficient fellow, to achieve similar outputs as their best performing peers. The model also identified the work intensity index as the key element that characterized the best performers in our fellowship. CONCLUSIONS: DEA is a feasible method of objectively evaluating peer performance in a critical care fellowship beyond summative evaluations alone and can potentially be a powerful tool to guide individual performance during the fellowship.

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Inadvertent Perioperative Hypothermia Induced by Spinal Anesthesia for Cesarean Delivery Might Be More Significant Than We Think: Are We Doing Enough to Warm Our Parturients?

No abstract available

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Society for Obstetric Anesthesia and Perinatology 2017 Meeting Report

imageNo abstract available

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Efficiency, Paths, Goals, and Frontiers in Graduate Education: New Uses for Old Concepts

imageNo abstract available

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Pentathol Postcards: Erratum

No abstract available

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Are You Down With TPP? Considering Transpulmonary Pressures as Opposed to Ventilator-Measured Pressures

No abstract available

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Repeated Administration of Duloxetine Suppresses Neuropathic Pain by Accumulating Effects of Noradrenaline in the Spinal Cord

imageBACKGROUND: Antidepressants are used to treat neuropathic pain and although the detailed mechanisms of their effects are unclear, the descending noradrenergic inhibitory system might play an important role. We tested our hypothesis that repeated administration of duloxetine suppresses neuropathic pain by restoring the descending noradrenergic inhibitory system in rats 6 weeks after spinal nerve ligation (SNL). METHODS: We subcutaneously injected SNL rats with duloxetine (10 mg kg−1 day−1) daily for 3 consecutive days and assessed behavioral hypersensitivity and noxious stimulus–induced analgesia (NSIA) activated by subcutaneous injection of capsaicin. We also performed microdialysis studies of the spinal cord, noradrenaline measurements of homogenized lumbar spinal tissue, and immunohistochemistry of the locus coeruleus. RESULTS: Three daily injections of duloxetine attenuated the mechanical hyperalgesia induced by SNL (SNL treated with vehicle: 88 ± 9.4 g versus SNL treated with duloxetine: 148 ± 13 g, P

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“However Beautiful the Strategy, You Should Occasionally Look at the Results”: Sir Winston Churchill and Medical Checklists

No abstract available

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Anaesthesia for the Elderly Patient, 2nd ed.

No abstract available

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Postoperative Atrial Fibrillation and Maslow’s Hammer

No abstract available

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In Response

No abstract available

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The Pediatric Elephant in the Room

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Risk of Cognitive Impairment by Sleep-Disordered Breathing

No abstract available

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The Eye of the Beholder

No abstract available

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Rehabilitation of Visual Loss: Where We Are and Where We Need to Be

Background: Spontaneous recovery of visual loss resulting from injury to the brain is variable. A variety of traditional rehabilitative strategies, including the use of prisms or compensatory saccadic eye movements, have been used successfully to improve visual function and quality-of-life for patients with homonymous hemianopia. More recently, repetitive visual stimulation of the blind area has been reported to be of benefit in expanding the field of vision. Evidence Acquisition: We performed a literature review with main focus on clinical studies spanning from 1963 to 2016, including 52 peer-reviewed articles, relevant cross-referenced citations, editorials, and reviews. Results: Repetitive visual stimulation is reported to expand the visual field, although the interpretation of results is confounded by a variety of methodological factors and conflicting outcomes from different research groups. Many studies used subjective assessments of vision and did not include a sufficient number of subjects or controls. Conclusions: The available clinical evidence does not strongly support claims of visual restoration using repetitive visual stimulation beyond the time that spontaneous visual recovery might occur. This lack of firm supportive evidence does not preclude the potential of real benefit demonstrated in laboratories. Additional well-designed clinical studies with adequate controls and methods to record ocular fixation are needed. Address correspondence to Behzad Mansouri, MD, PhD, FRCPC, Neurology Section, Department of Internal Medicine, Department of Ophthalmology, and Biomedical Engineering Program, University of Manitoba, 2735 Pembina Hwy, R3T2H5, Winnipeg, Manitoba, Canada; E-mail: behzad.mansouri@umanitoba.ca. The authors report no conflicts of interest. Supplemental digital content is available for this article. Direct URL citations appear in the printed text and are provided in the full text and PDF versions of this article on the journal's Web site (http://ift.tt/2BFTkP1). © 2017 by North American Neuro-Ophthalmology Society

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Optical Coherence Tomography in Optic Nerve Hypoplasia: Correlation With Optic Disc Diameter, Nerve Fiber Layer Thickness, and Visual Function

Background: The correlation between optic disc diameters (DDs) with average retinal nerve fiber layer thickness (RNFLT) and visual function in children with optic nerve hypoplasia (ONH) having nystagmus is unknown. Methods: Data were obtained from a retrospective review of 28 children (mean age: 9.4 years; ±5.1). Optic DD was defined as the maximal horizontal opening of Bruch membrane with spectral optical coherence tomography combined with a confocal laser ophthalmoscope. Average RNFLT was obtained from circumpapillary b-scans. RNFLT was also remeasured at eccentricities that were proportionate with DD to rule out potential sampling artifacts. Visual function was assessed by visual acuity at last follow-up and by visual evoked potentials (VEP) in 11 patients. The eye with the larger DD, which had better visual acuity, was analyzed to exclude potential effects of amblyopia. Results: DD was correlated with average RNFLT (r2 = 0.61), visual acuity (r2 = 0.32), and VEPs (r2 = 0.66). The relationship between RNFLT and DD was as follows: average RNFLT (μm) = 0.074 * DD (μm) − 18.8. RNFLT also correlated with the ratio of horizontal optic DD to macula-disc-margin distance (DD:DM; r2 = 0.59). RNFLT measured at eccentricities proportionate with DD showed progressive decrease in thickness only for DDs

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The Design and Assessment of a Multiparametric Model for the Dysphonia Severity Index for Persian-speaking Populations

In instrumental voice assessment, multiparametric models reflect the multidimensional nature of voice and are therefore better than models that reflect only a single dimension of voice. The Dysphonia Severity Index (DSI) is one of the most common multiparametric models. In voice assessment, race, language, and structural and physiological features affect the acoustic, aerodynamic, and voice range profile measures. Given these differences, this study was conducted to design and evaluate a multiparametric and objective model for assessing the severity of dysphonia in Persian-speaking populations.

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Modulation of the Alternative Pathway of Complement by Murine Factor H-Related Proteins [INNATE IMMUNITY AND INFLAMMATION]

Factor H (FH) is a key alternative pathway regulator that controls complement activation both in the fluid phase and on specific cell surfaces, thus allowing the innate immune response to discriminate between self and foreign pathogens. However, the interrelationships between FH and a group of closely related molecules, designated the FH-related (FHR) proteins, are currently not well understood. Whereas some studies have suggested that human FHR proteins possess complement regulatory abilities, recent studies have shown that FHR proteins are potent deregulators. Furthermore, the roles of the FHR proteins have not been explored in any in vivo models of inflammatory disease. In this study, we report the cloning and expression of recombinant mouse FH and three FHR proteins (FHR proteins A–C). Results from functional assays show that FHR-A and FHR-B proteins antagonize the protective function of FH in sheep erythrocyte hemolytic assays and increase cell-surface C3b deposition on a mouse kidney proximal tubular cell line (TEC) and a human retinal pigment epithelial cell line (ARPE-19). We also report apparent KD values for the binding interaction of mouse C3d with mouse FH (3.85 μM), FHR-A (136 nM), FHR-B (546 nM), and FHR-C (1.04 μM), which directly correlate with results from functional assays. Collectively, our work suggests that similar to their human counterparts, a subset of mouse FHR proteins have an important modulatory role in complement activation. Further work is warranted to define the in vivo context-dependent roles of these proteins and determine whether FHR proteins are suitable therapeutic targets for the treatment of complement-driven diseases.



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IL-33-Responsive Group 2 Innate Lymphoid Cells Are Regulated by Female Sex Hormones in the Uterus [INNATE IMMUNITY AND INFLAMMATION]

Group 2 innate lymphoid cells (ILC2s) reside in multiple organs in the body, where they play roles in immunity, tissue homeostasis, and metabolic regulation. However, little is known about the regulatory mechanisms of ILC2s in different organs. Here, we identified ILC2s in the mouse uterus and found that they express cell surface molecules, including the IL-33 receptor, ST2, that are roughly comparable to those expressed by lung ILC2s. Both in vivo and in vitro treatment with IL-33 induced type 2 cytokine production in uterine ILC2s, suggesting that they respond to IL-33 in a manner similar to ILC2s in other organs. Importantly, uterine ILC2s were nearly absent in ovariectomized mice and were increased in wild-type mice by estrogen administration, whereas lung ILC2s were unaffected by both ovariectomy and estrogen administration. Likewise, a marked reduction in uterine ILC2s was observed in mice deficient in estrogen receptor α or estrogen receptor β. Furthermore, uterine ILC2s highly expressed estrogen receptor α, and in vitro culture of isolated uterine ILC2s with 17β-estradiol modified expression of a number of genes. Finally, an increased prevalence in neonatal mortality was observed in litters from dams lacking the IL-33 receptor, ST2. Taken together, our findings indicate that unlike lung IL2Cs, uterine ILC2s are regulated by female sex hormones, which may specialize them for specific physiological functions.



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A Beginners Guide to Analyzing and Visualizing Mass Cytometry Data [IMMUNOLOGY NOTES AND RESOURCES]

Mass cytometry has revolutionized the study of cellular and phenotypic diversity, significantly expanding the number of phenotypic and functional characteristics that can be measured at the single-cell level. This high-dimensional analysis platform has necessitated the development of new data analysis approaches. Many of these algorithms circumvent traditional approaches used in flow cytometric analysis, fundamentally changing the way these data are analyzed and interpreted. For the beginner, however, the large number of algorithms that have been developed, as well as the lack of consensus on best practices for analyzing these data, raise multiple questions: Which algorithm is the best for analyzing a dataset? How do different algorithms compare? How can one move beyond data visualization to gain new biological insights? In this article, we describe our experiences as recent adopters of mass cytometry. By analyzing a single dataset using five cytometry by time-of-flight analysis platforms (viSNE, SPADE, X-shift, PhenoGraph, and Citrus), we identify important considerations and challenges that users should be aware of when using these different methods and common and unique insights that can be revealed by these different methods. By providing annotated workflow and figures, these analyses present a practical guide for investigators analyzing high-dimensional datasets. In total, these analyses emphasize the benefits of integrating multiple cytometry by time-of-flight analysis algorithms to gain complementary insights into these high-dimensional datasets.



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Toxoplasma gondii Inactivates Human Plasmacytoid Dendritic Cells by Functional Mimicry of IL-10 [INFECTIOUS DISEASE AND HOST RESPONSE]

Plasmacytoid dendritic cells (pDCs) are the major producers of IFN-α, an antiviral cytokine involved in immunomodulation and control of HIV type 1 replication, whereas Toxoplasma gondii is a life-threatening opportunistic infection in AIDS patients. During infection with HIV type 1, human pDCs decrease in circulation and remaining pDC produce lower amounts of IFN-α in response to viral stimulation. In this study, we investigated the impact of coinfection with T. gondii on the innate virus-directed responses of human pDCs. Using intracellular flow cytometry and fluorescence microscopy, we determined that T. gondii invaded but did not induce IFN-α or TNF-α in human pDC. However, T. gondii inhibited IFN-α and TNF-α produced in response to HSV and HIV, thus functionally inactivating pDC. IFN-α production was inhibited only in cells infected by T. gondii, which inhibited neither uptake of GFP-HSV nor localization of TLR9 in CD71+ endosomes, directing us to investigate downstream events. Using imaging flow cytometry, we found that both T. gondii and IL-10 inhibited virus-induced nuclear translocation, but not phosphorylation, of IFN response factor 7. Blockade of IFN response factor 7 nuclear translocation and inhibition of the IFN-α response was partially reversed by a deficiency in the T. gondii–derived ROP16 kinase, known to directly phosphorylate STAT3, a critical mediator of IL-10's anti-inflammatory effects. Taken together, our results indicate that T. gondii suppresses pDC activation by mimicking IL-10's regulatory effects through an ROP16 kinase-dependent mechanism. Our findings further imply a convergent mechanism of inhibition of TLR signaling by T. gondii and IL-10 and suggest potential negative consequences of HIV/T. gondii coinfection.



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The Reign of Antibodies: A Celebration of and Tribute to Michael Potter and His Homogeneous Immunoglobulin Workshops [PILLARS OF IMMUNOLOGY]



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Embryonic Lethality and Host Immunity of RelA-Deficient Mice Are Mediated by Both Apoptosis and Necroptosis [INNATE IMMUNITY AND INFLAMMATION]

In mammalian cells, signaling pathways triggered by TNF can be switched from NF-B activation to apoptosis and/or necroptosis. The in vivo mechanisms underlying the mutual regulation of these three signaling pathways are poorly understood. In this article, we report that the embryonic lethality of RelA-deficient mice is partially prevented by the deletion of Rip3 or Mlkl, but it is fully rescued by the combined ablation of Fadd and Rip3 or Mlkl or by blocking RIP1 kinase activity (RIP1K45A). RelA–/–Fadd–/–Rip3–/– triple-knockout (TKO) and RelA–/–Rip1K45A/K45A mice displayed bacterial pneumonia leading to death ~2 wk after birth. Moreover, RelA–/–Rip1K45A/K45A mice, but not TKO mice, developed severe inflammation associated with inflammatory skin lesion. Antibiotic treatment improved bacterial pneumonia, extended the lifespan of TKO and RelA–/–Rip1K45A/K45A mice, and alleviated skin inflammation in RelA–/–Rip1K45A/K45A mice. These results show the mechanisms underlying the in vivo mutual regulation between NF-B activation and the cell death pathway and provide new insights into this interplay in embryonic development and host immune homeostasis.



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Pillars Article: Evidence for Amino Acid Sequence Differences among Proteins Resembling the L-chain Subunits of Immunoglobulins. J. Mol. Biol. 1965. 12: 81-87 [PILLARS OF IMMUNOLOGY]



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In This Issue [IN THIS ISSUE]



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Salmonella Vaccines: Conduits for Protective Antigens [BRIEF REVIEWS]

Vaccines afford a better and more cost-effective approach to combatting infectious diseases than continued reliance on antibiotics or antiviral or antiparasite drugs in the current era of increasing incidences of diseases caused by drug-resistant pathogens. Recombinant attenuated Salmonella vaccines (RASVs) have been significantly improved to exhibit the same or better attributes than wild-type parental strains to colonize internal lymphoid tissues and persist there to serve as factories to continuously synthesize and deliver rAgs. Encoded by codon-optimized pathogen genes, Ags are selected to induce protective immunity to infection by that pathogen. After immunization through a mucosal surface, the RASV attributes maximize their abilities to elicit mucosal and systemic Ab responses and cell-mediated immune responses. This article summarizes many of the numerous innovative technologies and discoveries that have resulted in RASV platforms that will enable development of safe efficacious RASVs to protect animals and humans against a diversity of infectious disease agents.



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The IFN Response in Bats Displays Distinctive IFN-Stimulated Gene Expression Kinetics with Atypical RNASEL Induction [INFECTIOUS DISEASE AND HOST RESPONSE]

Bats host a large number of zoonotic viruses, including several viruses that are highly pathogenic to other mammals. The mechanisms underlying this rich viral diversity are unknown, but they may be linked to unique immunological features that allow bats to act as asymptomatic viral reservoirs. Vertebrates respond to viral infection by inducing IFNs, which trigger antiviral defenses through IFN-stimulated gene (ISG) expression. Although the IFN system of several bats is characterized at the genomic level, less is known about bat IFN-mediated transcriptional responses. In this article, we show that IFN signaling in bat cells from the black flying fox (Pteropus alecto) consists of conserved and unique ISG expression profiles. In IFN-stimulated cells, bat ISGs comprise two unique temporal subclusters with similar early induction kinetics but distinct late-phase declines. In contrast, human ISGs lack this decline phase and remained elevated for longer periods. Notably, in unstimulated cells, bat ISGs were expressed more highly than their human counterparts. We also found that the antiviral effector 2-5A–dependent endoribonuclease, which is not an ISG in humans, is highly IFN inducible in black flying fox cells and contributes to cell-intrinsic control of viral infection. These studies reveal distinctive innate immune features that may underlie a unique virus–host relationship in bats.



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The Role of MHC-E in T Cell Immunity Is Conserved among Humans, Rhesus Macaques, and Cynomolgus Macaques [ANTIGEN RECOGNITION AND RESPONSES]

MHC-E is a highly conserved nonclassical MHC class Ib molecule that predominantly binds and presents MHC class Ia leader sequence-derived peptides for NK cell regulation. However, MHC-E also binds pathogen-derived peptide Ags for presentation to CD8+ T cells. Given this role in adaptive immunity and its highly monomorphic nature in the human population, HLA-E is an attractive target for novel vaccine and immunotherapeutic modalities. Development of HLA-E–targeted therapies will require a physiologically relevant animal model that recapitulates HLA-E–restricted T cell biology. In this study, we investigated MHC-E immunobiology in two common nonhuman primate species, Indian-origin rhesus macaques (RM) and Mauritian-origin cynomolgus macaques (MCM). Compared to humans and MCM, RM expressed a greater number of MHC-E alleles at both the population and individual level. Despite this difference, human, RM, and MCM MHC-E molecules were expressed at similar levels across immune cell subsets, equivalently upregulated by viral pathogens, and bound and presented identical peptides to CD8+ T cells. Indeed, SIV-specific, Mamu-E–restricted CD8+ T cells from RM recognized antigenic peptides presented by all MHC-E molecules tested, including cross-species recognition of human and MCM SIV-infected CD4+ T cells. Thus, MHC-E is functionally conserved among humans, RM, and MCM, and both RM and MCM represent physiologically relevant animal models of HLA-E–restricted T cell immunobiology.



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The Activation of Human Dermal Microvascular Cells by Poly(I:C), Lipopolysaccharide, Imiquimod, and ODN2395 Is Mediated by the Fli1/FOXO3A Pathway [INNATE IMMUNITY AND INFLAMMATION]

Endothelial cell (EC) dysfunction has been associated with inflammatory and autoimmune diseases; however, the factors contributing to this dysfunction have not been fully explored. Because activation of TLRs has been implicated in autoimmune diseases, the goal of this study was to determine the effects of TLR ligands on EC function. Human dermal microvascular ECs (HDMECs) treated with TLR3 [Poly(I:C)], TLR4 (LPS), and TLR7 (imiquimod) agonists showed decreased proliferation and a reduced total number of branching tubules in three-dimensional human dermal organoid ex vivo culture. In contrast, the TLR9 ligand class C, ODN2395, increased angiogenesis. The antiproliferative effects of TLR3, TLR4, and TLR7 ligands correlated with significant downregulation of a key regulator of vascular homeostasis, Fli1, whereas TLR9 increased Fli1 levels. Furthermore, Poly(I:C) and LPS induced endothelial to mesenchymal transition that was reversed by the pretreatment with TGF-β neutralizing Ab or re-expression of Fli1. We showed that Fli1 was required for the HDMEC proliferation by transcriptionally repressing FOXO3A. In contrast to TLR9, which suppressed activation of the FOXO3A pathway, TLR3, TLR4, and TLR7 ligands activated FOXO3A as indicated by decreased phosphorylation and increased nuclear accumulation. The inverse correlation between Fli1 and FOXO3A was also observed in the vasculature of scleroderma patients. This work revealed opposing effects of TLR9 and TLR3, TLR4, and TLR7 on the key angiogenic pathways, Fli1 and FOXO3A. Our results provide a mechanistic insight into the regulation of angiogenesis by TLRs and confirm a central role of Fli1 in regulating vascular homeostasis.



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Superoxide Production by NADPH Oxidase Intensifies Macrophage Antiviral Responses during Diabetogenic Coxsackievirus Infection [AUTOIMMUNITY]

Coxsackievirus B infections are suspected environmental triggers of type 1 diabetes (T1D) and macrophage antiviral responses may provide a link to virus-induced T1D. We previously demonstrated an important role for NADPH oxidase (NOX)–derived superoxide production during T1D pathogenesis, as NOX-deficient NOD mice (NOD.Ncf1m1J) were protected against T1D due, in part, to impaired proinflammatory TLR signaling in NOD.Ncf1m1J macrophages. Therefore, we hypothesized that loss of NOX-derived superoxide would dampen diabetogenic antiviral macrophage responses and protect from virus-induced diabetes. Upon infection with a suspected diabetogenic virus, Coxsackievirus B3 (CB3), NOD.Ncf1m1J mice remained resistant to virus-induced autoimmune diabetes. A concomitant decrease in circulating inflammatory chemokines, blunted antiviral gene signature within the pancreas, and reduced proinflammatory M1 macrophage responses were observed. Importantly, exogenous superoxide addition to CB3-infected NOD.Ncf1m1J bone marrow–derived macrophages rescued the inflammatory antiviral M1 macrophage response, revealing reduction-oxidation–dependent mechanisms of signal transducer and activator of transcription 1 signaling and dsRNA viral sensors in macrophages. We report that superoxide production following CB3 infection may exacerbate pancreatic β cell destruction in T1D by influencing proinflammatory M1 macrophage responses, and mechanistically linking oxidative stress, inflammation, and diabetogenic virus infections.



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Granzyme A in Human Platelets Regulates the Synthesis of Proinflammatory Cytokines by Monocytes in Aging [INNATE IMMUNITY AND INFLAMMATION]

Dysregulated inflammation is implicated in the pathobiology of aging, yet platelet–leukocyte interactions and downstream cytokine synthesis in aging remains poorly understood. Platelets and monocytes were isolated from healthy younger (age <45, n = 37) and older (age ≥65, n = 30) adults and incubated together under autologous and nonautologous conditions. Synthesis of inflammatory cytokines by monocytes, alone or in the presence of platelets, was examined. Next-generation RNA-sequencing allowed for unbiased profiling of the platelet transcriptome in aging. Basal IL-8 and MCP-1 synthesis by monocytes alone did not differ between older and younger adults. However, in the presence of autologous platelets, monocytes from older adults synthesized greater IL-8 (41 ± 5 versus 9 ± 2 ng/ml, p < 0.0001) and MCP-1 (867 ± 150 versus 216 ± 36 ng/ml, p < 0.0001) than younger adults. Platelets from older adults were sufficient for upregulating the synthesis of inflammatory cytokines by monocytes. Using RNA-sequencing of platelets followed by validation via RT-PCR and immunoblot, we discovered that granzyme A (GrmA), a serine protease not previously identified in human platelets, increases with aging (~9-fold versus younger adults, p < 0.05) and governs increased IL-8 and MCP-1 synthesis through TLR4 and caspase-1. Inhibiting GrmA reduced excessive IL-8 and MCP-1 synthesis in aging to levels similar to younger adults. In summary, human aging is associated with changes in the platelet transcriptome and proteome. GrmA is present and bioactive in human platelets, is higher in older adults, and controls the synthesis of inflammatory cytokines by monocytes. Alterations in the platelet molecular signature and signaling to monocytes may contribute to dysregulated inflammatory syndromes in older adults.



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Essential Role of CARD14 in Murine Experimental Psoriasis [AUTOIMMUNITY]

Caspase recruitment domain family member 14 (CARD14) was recently identified as a psoriasis-susceptibility gene, but its immunological role in the pathogenesis of psoriasis in vivo remains unclear. In this study, we examined the role of CARD14 in murine experimental models of psoriasis induced by either imiquimod (IMQ) cream or recombinant IL-23 injection. In all models tested, the psoriasiform skin inflammation was abrogated in Card14–/– mice. Comparison of the early gene signature of the skin between IMQ-cream–treated Card14–/– mice and Tlr7–/–Tlr9–/– mice revealed not only their similarity, but also distinct gene sets targeted by IL-23. Cell type–specific analysis of these mice identified skin Langerinhigh Langerhans cells as a potent producer of IL-23, which was dependent on both TLR7 and TLR9 but independent of CARD14, suggesting that CARD14 is acting downstream of IL-23, not TLR7 or TLR9. Instead, a bone marrow chimera study suggested that CARD14 in radio-sensitive hematopoietic cells was required for IMQ-induced psoriasiform skin inflammation, controlling the number of V4+ T cells producing IL-17 or IL-22 infiltrating through the dermis to the inflamed epidermis. These data indicate that CARD14 is essential and a potential therapeutic target for psoriasis.



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Identity and Diversity of Human Peripheral Th and T Regulatory Cells Defined by Single-Cell Mass Cytometry [SYSTEMS IMMUNOLOGY]

Human CD3+CD4+ Th cells, FOXP3+ T regulatory (Treg) cells, and T regulatory type 1 (Tr1) cells are essential for ensuring peripheral immune response and tolerance, but the diversity of Th, Treg, and Tr1 cell subsets has not been fully characterized. Independent functional characterization of human Th1, Th2, Th17, T follicular helper (Tfh), Treg, and Tr1 cells has helped to define unique surface molecules, transcription factors, and signaling profiles for each subset. However, the adequacy of these markers to recapitulate the whole CD3+CD4+ T cell compartment remains questionable. In this study, we examined CD3+CD4+ T cell populations by single-cell mass cytometry. We characterize the CD3+CD4+ Th, Treg, and Tr1 cell populations simultaneously across 23 memory T cell–associated surface and intracellular molecules. High-dimensional analysis identified several new subsets, in addition to the already defined CD3+CD4+ Th, Treg, and Tr1 cell populations, for a total of 11 Th cell, 4 Treg, and 1 Tr1 cell subsets. Some of these subsets share markers previously thought to be selective for Treg, Th1, Th2, Th17, and Tfh cells, including CD194 (CCR4)+FOXP3+ Treg and CD183 (CXCR3)+T-bet+ Th17 cell subsets. Unsupervised clustering displayed a phenotypic organization of CD3+CD4+ T cells that confirmed their diversity but showed interrelation between the different subsets, including similarity between Th1–Th2–Tfh cell populations and Th17 cells, as well as similarity of Th2 cells with Treg cells. In conclusion, the use of single-cell mass cytometry provides a systems-level characterization of CD3+CD4+ T cells in healthy human blood, which represents an important baseline reference to investigate abnormalities of different subsets in immune-mediated pathologies.



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The Loss of TET2 Promotes CD8+ T Cell Memory Differentiation [IMMUNE REGULATION]

T cell differentiation requires appropriate regulation of DNA methylation. In this article, we demonstrate that the methylcytosine dioxygenase ten-eleven translocation (TET)2 regulates CD8+ T cell differentiation. In a murine model of acute viral infection, TET2 loss promotes early acquisition of a memory CD8+ T cell fate in a cell-intrinsic manner without disrupting Ag-driven cell expansion or effector function. Upon secondary recall, TET2-deficient memory CD8+ T cells demonstrate superior pathogen control. Genome-wide methylation analysis identified a number of differentially methylated regions in TET2-deficient versus wild-type CD8+ T cells. These differentially methylated regions did not occur at the loci of differentially expressed memory markers; rather, several hypermethylated regions were identified in known transcriptional regulators of CD8+ T cell memory fate. Together, these data demonstrate that TET2 is an important regulator of CD8+ T cell fate decisions.



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